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JXB Advance Access originally published online on May 19, 2009
Journal of Experimental Botany 2009 60(11):3179-3193; doi:10.1093/jxb/erp153
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© The Author [2009]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

AtRAB-H1b and AtRAB-H1c GTPases, homologues of the yeast Ypt6, target reporter proteins to the Golgi when expressed in Nicotiana tabacum and Arabidopsis thaliana

Jorunn Nergaard Johansen1 *, Cheung-Ming Chow2 {dagger}, Ian Moore2 and Chris Hawes1,{ddagger}

1School of Life Sciences, Oxford Brookes University, Gipsy Lane, Oxford OX3 0BP, UK
2Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, UK

{ddagger} To whom correspondence should be addressed: E-mail: chawes{at}brookes.ac.uk

Ypt/Rab GTPases act as key regulators of intracellular traffic through the conformational differences exhibited by their GTP or GDP-bound forms. In this paper, two Arabidopsis Ypt6 homologues, AtRAB-H1b and AtRAB-H1c were characterized and compared. Using a live cell imaging approach, it is shown that yellow fluorescent protein-fusions (YFP) of AtRAB-H1b and AtRAB-H1c locate to the Golgi and to the cytosol in both Nicotiana tabacum and in Arabidopsis thaliana. In addition, YFP-AtRAB-H1b targets an as yet unknown compartment not labelled by YFP-AtRAB-H1c or Golgi markers. It is also shown that the subcellular location of YFP-AtRAB-H1b and YFP-AtRAB-H1c is affected by the state of GTP-binding and that expression of a GTP-deficient mutant results in increased apoplastic fluorescence of a secretory form of YFP.

Key words: Confocal microscopy, GFP, Golgi apparatus, Rab GTPase

* Present address: Institute for Cancer Research, Department of Immunology, Rikshospitalet-Radiumhospitalet Medical Centre, Montebello 0310 Oslo, Norway.

{dagger} Department of Biology, Science Centre, The Chinese University of Hong Kong, Shatin, Hong Kong SAR.

Received 16 December 2008; Revised 17 April 2009 Accepted 20 April 2009


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